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Team:Washington/Celiacs/Future
From 2011.igem.org
Kinetic Characterization
We intend to develop a mass spectroscopy assay to measure the kcat and kM values for our best mutant.
Biophysical Characterization
Once the ideal mutations are isolated, we intend to test the best mutants at gastric pH and in the presence of other gastric enzymes for a short period of time, mimicking the environment after enzyme ingestion and prior to enzyme uptake by the small intestine. We suspect that the thermostable properties of Kumamolisin-As will render our mutant enzyme reasonably resistant to degradation by gastric enzymes such as pepsin. However, if this proves not to be the case, we intend to reengineer the mutant for enhanced resistance to pepsin and other such gastric enzymes. Once ensured that the mutated Kumamolisin-As remains active under stomach conditions, this ideal mutated enzyme will be ready for in vivo experimentation.
